Use of adenine base editing and homology-independent targeted integration strategies to correct the cystic fibrosis causing variant, W1282X.

Small molecule drugs known as modulators can treat ~90% of people with cystic fibrosis (CF), but do not work for premature termination codon variants such as W1282X (c.3846G>A). Here we evaluated two gene editing strategies, Adenine Base Editing (ABE) to correct W1282X, and Homology-Independent Targeted Integration (HITI) of a CFTR superexon comprising exons 23-27 (SE23-27) to enable expression of a CFTR mRNA without W1282X. In Flp-In-293 cells stably expressing a CFTR expression minigene bearing W1282X, ABE corrected 24% of W1282X alleles, rescued CFTR mRNA from nonsense mediated decay and restored protein expression. However, bystander editing at the adjacent adenine (c.3847A>G), caused an amino acid change (R1283G) that affects CFTR maturation and ablates ion channel activity. In primary human nasal epithelial cells homozygous for W1282X, ABE corrected 27% of alleles, but with a notably lower level of bystander editing, and CFTR channel function was restored to 16% of wild-type levels. Using the HITI approach, correct integration of a SE23-27 in intron 22 of the CFTR locus in 16HBEge W1282X cells was detected in 5.8% of alleles, resulting in 7.8% of CFTR transcripts containing the SE23-27 sequence. Analysis of a clonal line homozygous for the HITI-SE23-27 produced full-length mature protein and restored CFTR anion channel activity to 10% of wild-type levels, which could be increased three-fold upon treatment with the triple combination of CF modulators. Overall, these data demonstrate two different editing strategies can successfully correct W1282X, the second most common class I variant, with a concomitant restoration of CFTR function.

Evaluation of the potential of achachairu peel (Garcinia humilis) for the fortification of cereal-based foods.

BACKGROUND: Achachairu is an exotic fruit that is being studied for its bioactive compound composition. However, there is scarce information on the properties of its by-products and their incorporation into food. In this study, achachairu peels were used to obtain phenolic-rich and sustainable ingredients. Furthermore, their potential for use in the fortification of food, particularly cereal-based products, was evaluated. Different ratios of ethanol:water were used to extract the phenolic compounds from achachairu (100:0, 90:10 and 70:30). The optimal extract was characterised regarding its total phenolic content (TPC) and antioxidant, antimicrobial and anti-enzymatic properties. Finally, achachairu peel powder and extract were incorporated into bread and their influence on the texture, antioxidant properties and shelf life of the product was evaluated. RESULTS: High-performance liquid chromatographic analysis showed that the 90:10 extract contained a higher phenolic composition than the other samples. The achachairu extract presented a TPC of 88.7 mgGAE g-1, good antioxidant capacity towards DPPH and ABTS radicals and the capacity to inhibit the activity of α-amylase by almost 80%. The addition of achachairu peel powder and extract to bread increased its hardness, chewiness and gumminess, not affecting the remaining texture parameters. An increase in the TPC, antioxidant properties and shelf life of the product was also observed. CONCLUSION: This study proves the potential for achachairu by-products incorporated into cereal-based products to improve their biological properties while extending the food shelf life. © 2024 Society of Chemical Industry.

Ferrozoles: Ferrocenyl derivatives of letrozole with dual effects as potent aromatase inhibitors and cytostatic agents.

In the treatment of hormone-dependent cancers, aromatase inhibitors (AI) are receiving increased attention due to some undesirable effects such as the risk of endometrial cancer and thromboembolism of SERMs (selective estrogen receptor modulators). Letrozole is the most active AI with 99% aromatase inhibition. Unfortunately, this compound also exhibits some adverse effects such as hot flashes and fibromyalgias. Therefore, there is an urgent need to explore new types of AIs that retain the same-or even increased-antitumor ability. Inspired by the letrozole structure, a set of new derivatives has been synthesized that include a ferrocenyl moiety and different heterocycles. The derivative that contains a benzimidazole ring, namely compound 6, exhibits a higher aromatase inhibitory activity than letrozole and it also shows potent cytostatic behavior when compared to other well-established aromatase inhibitors, as demonstrated by dose-response, cell cycle, apoptosis and time course experiments. Furthermore, 6 promotes the inhibition of cell growth in both an aromatase-dependent and -independent fashion, as indicated by the study of A549 and MCF7 cell lines. Molecular docking and molecular dynamics calculations on the interaction of 6 or letrozole with the aromatase binding site revealed that the ferrocene moiety increases the van der Waals and hydrophobic interactions, thus resulting in an increase in binding affinity. Furthermore, the iron atom of the ferrocene fragment can form a metal-acceptor interaction with a propionate fragment, and this results in a stronger coupling with the heme group-a possibility that is consistent with the strong aromatase inhibition of 6.

Sucrose analgesia for heel-lance procedures in neonates.

BACKGROUND: Sucrose has been examined for calming and pain-relieving effects in neonates for invasive procedures such as heel lance. OBJECTIVES: To assess the effectiveness of sucrose for relieving pain from heel lance in neonates in terms of immediate and long-term outcomes SEARCH METHODS: We searched (February 2022): CENTRAL, MEDLINE, Embase, PsycINFO, CINAHL, Web of Science, and three trial registries. SELECTION CRITERIA: We included randomised controlled trials where term and/or preterm neonates received sucrose for heel lances. Comparison treatments included water/placebo/no intervention, non-nutritive sucking (NNS), glucose, breastfeeding, breast milk, music, acupuncture, facilitated tucking, and skin-to-skin care. DATA COLLECTION AND ANALYSIS: We used standard Cochrane methods. We reported mean differences (MD) with 95% confidence intervals (CI) using the fixed-effect model for continuous outcome measures. We assessed heterogeneity by the I2 test. We used GRADE to assess certainty of evidence. MAIN RESULTS: We included 55 trials (6273 infants): 29 included term neonates, 22 included preterm neonates, and four included both. Heel lance was investigated in 50 trials; 15 investigated other minor painful procedures in addition to lancing. Sucrose vs control The evidence suggests that sucrose probably results in a reduction in PIPP scores compared to the control group at 30 seconds (MD -1.74 (95% CI -2.11 to -1.37); I2 = 62%; moderate-certainty evidence) and 60 seconds after lancing (MD -2.14, 95% CI -3.34 to -0.94; I2 = 0%; moderate-certainty evidence). The evidence is very uncertain about the effects of sucrose on DAN scores compared to water at 30 seconds after lancing (MD -1.90, 95% CI -8.58 to 4.78; heterogeneity not applicable (N/A); very low-certainty evidence). The evidence suggests that sucrose probably results in a reduction in NIPS scores compared to water immediately after lancing (MD -2.00, 95% CI -2.42 to -1.58; heterogeneity N/A; moderate-certainty evidence). Sucrose vs NNS The evidence is very uncertain about the effect of sucrose on PIPP scores compared to NNS during the recovery period after lancing (MD 0.60, 95% CI -0.30 to 1.50; heterogeneity not applicable; very low-certainty evidence) and on DAN scores at 30 seconds after lancing (MD -1.20, 95% CI -7.87 to 5.47; heterogeneity N/A; very low-certainty evidence). Sucrose + NNS vs NNS The evidence is very uncertain about the effect of sucrose + NNS on PIPP scores compared to NNS during lancing (MD -4.90, 95% CI -5.73 to -4.07; heterogeneity not applicable; very low-certainty evidence) and during recovery after lancing (MD -3.80, 95% CI -4.47 to -3.13; heterogeneity N/A; very low-certainty evidence). The evidence is very uncertain about the effects of sucrose + NNS on NFCS scores compared to water + NNS during lancing (MD -0.60, 95% CI -1.47 to 0.27; heterogeneity N/A; very low-certainty evidence). Sucrose vs glucose The evidence suggests that sucrose results in little to no difference in PIPP scores compared to glucose at 30 seconds (MD 0.26, 95% CI -0.70 to 1.22; heterogeneity not applicable; low-certainty evidence) and 60 seconds after lancing (MD -0.02, 95% CI -0.79 to 0.75; heterogeneity N/A; low-certainty evidence). Sucrose vs breastfeeding The evidence is very uncertain about the effect of sucrose on PIPP scores compared to breastfeeding at 30 seconds after lancing (MD -0.70, 95% CI -0.49 to 1.88; I2 = 94%; very low-certainty evidence). The evidence is very uncertain about the effect of sucrose on COMFORTneo scores compared to breastfeeding after lancing (MD -2.60, 95% CI -3.06 to -2.14; heterogeneity N/A; very low-certainty evidence). Sucrose vs expressed breast milk The evidence suggests that sucrose may result in little to no difference in PIPP-R scores compared to expressed breast milk during (MD 0.3, 95% CI -0.24 to 0.84; heterogeneity not applicable; low-certainty evidence) and at 30 seconds after lancing (MD 0.3, 95% CI -0.11 to 0.71; heterogeneity N/A; low-certainty evidence). The evidence suggests that sucrose probably may result in slightly increased PIPP-R scores compared to expressed breast milk 60 seconds after lancing (MD 1.10, 95% CI 0.34 to 1.86; heterogeneity N/A; low-certainty evidence). The evidence is very uncertain about the effect of sucrose on DAN scores compared to expressed breast milk 30 seconds after lancing (MD -1.80, 95% CI -8.47 to 4.87; heterogeneity N/A; very low-certainty evidence). Sucrose vs laser acupuncture There was no difference in PIPP-R scores between sucrose and music groups; however, data were reported as medians and IQRs. The evidence is very uncertain about the effect of sucrose on NIPS scores compared to laser acupuncture during lancing (MD -0.86, 95% CI -1.43 to -0.29; heterogeneity N/A; very low-certainty evidence). Sucrose vs facilitated tucking The evidence is very uncertain about the effect of sucrose on total BPSN scores compared to facilitated tucking during lancing (MD -2.27, 95% CI -4.66 to 0.12; heterogeneity N/A; very low-certainty evidence) and during recovery after lancing (MD -0.31, 95% CI -1.72 to 1.10; heterogeneity N/A; very low-certainty evidence). Sucrose vs skin-to-skin + water (repeated lancing) The evidence suggests that sucrose results in little to no difference in PIPP scores compared to skin-to-skin + water at 30 seconds after 1st (MD 0.13, 95% CI -0.70 to 0.96); 2nd (MD -0.56, 95% CI -1.57 to 0.45); or 3rd lancing (MD-0.15, 95% CI -1.26 to 0.96); heterogeneity N/A, low-certainty evidence for all comparisons. The evidence suggests that sucrose results in little to no difference in PIPP scores compared to skin-to-skin + water at 60 seconds after 1st (MD -0.61, 95% CI -1.55 to 0.33); 2nd (MD -0.12, 95% CI -0.99 to 0.75); or 3rd lancing (MD-0.40, 95% CI -1.48 to 0.68); heterogeneity N/A, low-certainty evidence for all comparisons. Minor adverse events required no intervention. AUTHORS' CONCLUSIONS: Sucrose compared to control probably results in a reduction of PIPP scores 30 and 60 seconds after single heel lances (moderate-certainty evidence). Evidence is very uncertain about the effect of sucrose compared to NNS, breastfeeding, laser acupuncture, facilitated tucking, and the effect of sucrose + NNS compared to NNS in reducing pain. Sucrose compared to glucose, expressed breast milk, and skin-to-skin care shows little to no difference in pain scores. Sucrose combined with other nonpharmacologic interventions should be used with caution, given the uncertainty of evidence.

Innovative Approaches for Food: Using Natural Phenolic-Rich Extracts to Produce Value-Added Fresh Pasta.

Cereal-based products, which are rich in carbohydrates, are widely consumed worldwide; however, this type of food lacks other nutrients. Phenolic compounds from natural sources, such as Moringa oleifera, can be incorporated into these products to increase their nutritional and biological value. In this study, a phenolic-rich extract was obtained from M. oleifera leaf powder using a Soxhlet extractor. The extract obtained presented a total phenolic content of 79.0 mg of gallic acid equivalents/g and the ABTS and DPPH assays showed that the extract can act as an anti-oxidant agent, with IC50 values of 205.2 mg/L and 636.0 mg/L, respectively. Afterwards, fresh pasta was produced and the extract was incorporated into the pasta to improve its biological properties and extend its shelf-life. The results demonstrated that the addition of M. oleifera to the fresh pasta increased its anti-oxidant capacity and did not interfere with the cooking properties of the product. Moreover, the fortified pasta presented an increased shelf-life, since the extract conferred protection against microbial contamination for longer periods of time. Therefore, these findings showed that the incorporation of phenolic-rich extracts from natural sources (such as M. oleifera) is a feasible sustainable biotechnological approach to produce value-added cereal-based products.

A Comparative Study between Onion Peel Extracts, Free and Complexed with ß-Cyclodextrin, as a Natural UV Filter to Cosmetic Formulations.

The growing concern regarding the adverse effects of synthetic UV filters found in sunscreens has spurred significant attention due to their potential harm to aquatic ecosystems and human health. To address this, the present study aimed to extract and microencapsulate sensitive bioactive compounds derived from by-product onion peel (OP) by molecular inclusion using ß-cyclodextrin as the wall material. Identification and quantification of bioactive compounds within the extract were conducted through high-performance liquid chromatography (HPLC-DAD) analysis, revealing quercetin and resveratrol as the primary constituents. The photoprotection capacity, evaluated by the sun protection factor (SPF), revealed a protection factor comparable to the value for a synthetic UV filter. The produced microparticles presented high antioxidant capacity, significant photoprotection capacity, encapsulation efficiency of 91.8%, mean diameter of 31 µm, and polydispersity of 2.09. Furthermore, to comprehensively evaluate the performance of OP extract and its potential as a natural UV filter, five O/W emulsions were produced. Results demonstrated that microparticles displayed superior ability in maintaining SPF values over a five-week period. Photoprotection evaluation-skin reactivity tests revealed that both extract and microparticles absorb UV radiation in other regions of UV radiation, revealing their potential to be used as a natural UV filter to produce a sustainable and eco-friendly value-added sunscreen.

A Novel Approach in Skin Care: By-Product Extracts as Natural UV Filters and an Alternative to Synthetic Ones.

The cosmetic industry has been focusing on replacing synthetic ingredients with natural ones, taking advantage of their bioactive compounds. This work assessed the biological properties of onion peel (OP) and passion fruit peel (PFP) extracts in topical formulations as an alternative to synthetic antioxidants and UV filters. The extracts were characterized regarding their antioxidant capacity, antibacterial capacity and sun protection factor (SPF) value. Results revealed that the OP extract exhibited better results, which can result from the high concentrations of quercetin, as identified and quantified in HPLC analysis. Afterward, nine formulations of O/W creams were produced with minor changes in the quantity of additives: OP and PFP extract (natural antioxidants and UV filters), BHT (synthetic antioxidant) and oxybenzone (synthetic UV filter). The stability of the formulations was determined for 28 days; it was verified that they remained stable throughout the study period. The assays of the formulations' antioxidant capacity and SPF value revealed that OP and PFP extracts have some photoprotective properties and are excellent sources of antioxidants. As a result, they can be incorporated in daily moisturizers with SPF and sunscreens replacing and/or diminishing the quantities of synthetic ingredients, reducing their negative effects on human health and the environment.

Incorporation of Moringa oleifera Leaf Extract in Yoghurts to Mitigate Children's Malnutrition in Developing Countries.

Moringa oleifera, which is rich in bioactive compounds, has numerous biological activities and is a powerful source of antioxidants and nutrients. Therefore, M. oleifera can be incorporated into food to mitigate children's malnutrition. In this work, the bioactive compounds were extracted from M. oleifera leaf powder by ultrasound-assisted solid-liquid extraction. The antioxidant and antimicrobial activities and the phenolic composition of the extract were evaluated. The extract presented a total phenolic content of 54.5 ± 16.8 mg gallic acid equivalents/g and IC50 values of 133.4 ± 12.3 mg/L for DPPH and 60.0 ± 9.9 mg/L for ABTS. Catechin, chlorogenic acid, and epicatechin were the main phenolics identified by HPLC-DAD. The obtained extract and M. oleifera leaf powder were incorporated into yoghurts and their physicochemical and biological properties were studied. The incorporation of M. oleifera did not impair the yoghurts' stability over eight weeks when compared to both negative and positive controls. The extract presented higher stability regarding syneresis but lower stability regarding TPC compared to the powder. Also, the fortified yoghurts presented higher antioxidant properties than the negative control. These findings highlight the potential use of M. oleifera powder and extract as natural additives to produce fortified foods that can be used in the mitigation of malnutrition.

Physical immobilization of particles inspired by pollination.

Biomimetic systems often exhibit striking designs well adapted to specific functions that have been inspiring the development of new technologies. Herein, we explored the remarkable ability of honey bees to catch and release large quantities of pollen grains. Hair spacing and height on bees are crucial for their ability to mechanically fix pollen grains. Inspired by this, we proposed the concept of a micropatterned surface for microparticle entrapment, featuring high-aspect-ratio elastic micropillars spaced to mimic the hairy surface of bees. The hypothesis was validated by investigating the ability of polydimethylsiloxane microfabricated patches to fix microparticles. The geometrical arrangement, spacing, height, and flexibility of the fabricated micropillars, and the diameter of the microparticles, were investigated. Higher entrapment capability was found through the match between particle size and pillar spacing, being consistent with the observations that the diameter of pollen grains is similar to the spacing between hairs on bees' legs. Taller pillars permitted immobilization of higher quantities of particles, consistent with the high aspect ratio of bees' hairs. Our biomimetic surfaces were explored for their ability to fix solid microparticles for drug-release applications, using tetracycline hydrochloride as a model antibiotic. These surfaces allowed fixation of more than 20 mg/cm2 of antibiotic, about five times higher dose than commercialized patches (5.1 mg/cm2). Such bioinspired hairy surfaces could find applications in a variety of fields where dry fixation of high quantities of micrometer-sized objects are needed, including biomedicine, agriculture, biotechnology/chemical industry, and cleaning utensils.

A Potential Valorization Strategy of Wine Industry by-Products and Their Application in Cosmetics-Case Study: Grape Pomace and Grapeseed.

Grape pomace and grapeseed are agro-industrial by-products, whose inadequate treatment generates socioeconomic and environmental concerns. Nevertheless, it is possible to valorize them by extracting their bioactive compounds, such as antioxidants (phenolic compounds), vitamin E and fatty acids. The bioactive compounds were extracted using solid-liquid extraction. The yields for phenolic compounds were 18.4 ± 0.4% for grape pomace, and 17.4 ± 0.4%, for grapeseed. For the oil, the yields were 13.3 ± 0.2% and 14.5 ± 0.3% for grape pomace and grapeseed. Antioxidant capacity was assessed by the assay with 2,2-diphenyl-1-picrylhydrazyl (DPPH), and showed that phenolic extract has higher antioxidant capacity than the oils. Grape pomace and grapeseed extracts exhibit, correspondingly, values of 90.8 ± 0.8 and 87.5 ± 0.5 of DPPH inhibition and IC50 of 48.9 ± 0.5 and 55.9 ± 0.7 µgextract·mLDPPH-1. The antimicrobial capacity was assessed by the disk diffusion test, and revealed that, phenolic extracts inhibit the growth of Staphylococcus aureus and Staphylococcus epidermidis. The obtained extracts were incorporated in 10 face cream formulations, with slight modifications in quantities of formulation stabilizers. Their stability was studied for 35 days, and this revealed the possibility of incorporating extracts and oils obtained from by-products as antioxidants in cosmetics, and replacing synthetic ones. As a future recommendation, microencapsulation of the extracts should be performed, in order to increase their stability.

Response Surface Modeling and Optimization of the Extraction of Phenolic Antioxidants from Olive Mill Pomace.

Bioactive compounds from olive mill pomace (OMP) were extracted through a two-step solid-liquid extraction procedure considering four factors at five levels of a central composite rotatable response surface design. The influence of the process variables time of the primary extraction (2.0-4.0 h), solvent-to-sample ratio during the primary extraction (5.0-10.0 mL/g), time of the secondary extraction (1.0-2.0 h), and the solvent-to-sample ratio during the secondary extraction (3.0-5.0 mL/g) were examined. The content of bioactive compounds was determined spectrophotometrically, and the individual phenolic compounds were evaluated by reserved-phase high-performance liquid chromatography (RP-HPLC). The Derringer's function was used to optimize the extraction process, and the best conditions were found to be 3.2 h for the primary extraction, 10.0 mL/g for the solvent-to-sample ratio and 1.3 h for the secondary extraction associated with a solvent-to-sample ratio of 3.0 mL/g, obtaining a total phenolic content of 50.0 (expressed as mg gallic acid equivalents (GAE)/g dry weight (dw). The response surface methodology proved to be a great alternative for reducing the number of tests, allowing the optimization of the extraction of phenolic antioxidants from OMP with a reduced number of experiments, promoting reductions in cost and analysis time.

Sustainability in Skin Care: Incorporation of Avocado Peel Extracts in Topical Formulations.

The avocado peel is an agro-industrial by-product that has exhibited a massive increase in its production in the last few years. The reuse and valorisation of this by-product are essential since its disposal raises environmental concerns. In the present study, ethanolic extracts of avocado peels of the Hass variety were obtained, for three extraction times (1.5 h, 3 h and 4 h) and analysed for their antioxidant and antibacterial properties. Antioxidant evaluations of the extracts revealed that the extraction time of 1.5 h exhibited the best results amongst the three, with a DPPH inhibition percentage of 93.92 ± 1.29 and an IC50 percentage, the necessary concentration of the extract to inhibit 50% of DPPH, of 37.30 ± 1.00. The antibacterial capacity of the extracts was evaluated and it was revealed that they were able to inhibit the growth and development of bacteria of the Staphylococcus family. The obtained extract was incorporated in two types of cosmetic formulations (oil-in-water and water-in-oil) and their stability was evaluated and compared with formulations containing synthetic preservatives (BHT and phenoxyethanol). The results of the stability evaluation suggest that the avocado peel extract has the potential to be incorporated in both types of emulsions, acting as an antioxidant and antibacterial agent, proving it to be a viable option to reduce/replace the use of synthetic preservatives. Furthermore, the avocado peel extract proved to be more effective and stable in oil-in-water emulsions. These results highlight the possibility of obtaining sustainable cosmetics, significantly reducing the negative impacts on the environment by the incorporation of extracts sourced from the avocado peel, an interesting source of phenolic compounds, an abundant and low-cost by-product.

Allopurinol for fibromyalgia pain in adults: A randomized controlled trial.

BACKGROUND: Allopurinol is a potent inhibitor of the enzyme xanthine oxidase used in the treatment of hyperuricemia and gout. Because it is well known that purines exert multiple affects on pain transmission, we hypothesized that the inhibition of xanthine oxidase by allopurinol could be a valid strategy to treat pain in humans. This study aimed to compare the analgesic efficacy of oral allopurinol versus placebo as an adjuvant therapy in patients displaying fibromyalgia. METHODS: This randomized, double-blinded, placebo-controlled study included 60 women with the diagnosis of fibromyalgia. Patients were randomly assigned to receive either oral allopurinol 300 mg (n = 31) or placebo (n = 29) twice daily during 30 days. The patients were submitted to evaluation for pain sensitivity, anxiety, depression, and functional status before treatment, and 15 and 30 days thereafter. RESULTS: Oral administration of allopurinol 300 mg twice daily was ineffective in improving pain scores measured by several tools up to 30 days of treatment (P > 0.05). Additionally, no significant effects of allopurinol over anxiety, depressive symptoms, and functional status of fibromyalgia patients were observed in the present study. CONCLUSIONS: Although previous findings indicated that allopurinol could present intrinsic analgesic effects in both animals and humans, this study showed no benefit of the use of oral allopurinol as an adjuvant strategy during 30 days in women displaying fibromyalgia. However, considering previous promising results, new prospective studies are still valid to further investigate allopurinol and more selective purine derivatives in the management of pain syndromes.

Deriving valorization of phenolic compounds from olive oil by-products for food applications through microencapsulation approaches: a comprehensive review.

Nowadays, olive oil consumption is correlated to many health benefits, essentially due to the presence of antioxidants, especially phenolic compounds, which fostered its intensive production worldwide. During olive oil extraction, through continuous or discontinuous processes, many olive oil by-products are generated. These by-products constitute an environmental problem regarding its management and disposal. They are phytotoxic and biotoxic due to their high content of phenolic compounds, presenting contrastingly relevant health benefits due to their potent radical scavenging activities. In the framework of the disposal and management of olive oil by-products, treatment, and valorization approaches are found. As currently, the majority of the valorization techniques applied have a null market value, alternative strategies for the obtainment of innovative products as fortified foods are being investigated. The recovery and valorization strategies of olive oil by-products may comprise extraction and further encapsulation of bioactive compounds, as an innovative valorization blueprint of phenolic compounds present in these by-products. The majority of phenolic compounds present in olive oil by-products possess limited application on the food industry since they are promptly amended by environmental factors like temperature, pH, and light. Consequently, they must be protected previously ending in the final formulation. Prior to foods fortification with phenolic-rich extracts obtained from olive oil by-products, they should be protected through microencapsulation approaches, allowing a sustained release of phenolic compounds in the fortified foods, without losing their physicochemical properties. The combined strategies of extraction and microencapsulation will contribute to promoting the sustainability of the olive oil sector and aid the food industry to obtain reinvented added-value products.

Modified dispersive solid-phase extraction and cleanup followed by GC-MS/MS analysis to quantify ultraviolet filters and synthetic musk compounds in soil samples.

A simple method for the analysis of 13 synthetic musk compounds and six ultraviolet filters in soil samples was developed using a modified dispersive solid-phase methodology known as "Quick, Easy, Cheap, Effective, Rugged and Safe," followed by gas chromatography-triple quadrupole mass spectrometry. The methodology was validated by assessing linearity ranges, detection limits, precision, and accuracy. The method detection limit ranged between 0.01 and 10.00 ng/g dry weight and accuracy from 81 to 122%. A good precision was achieved, with relative standard deviation <10%. The applicability of the methodology was tested using different types of soils. Both synthetic musks and ultraviolet filters were detected in all soil samples. The most frequently detected compounds were benzophenone, octocrylene, 2-ethylhexyl 4-dimethylaminobenzoate, 2-ethylhexyl 4-methoxycinnamate, and galaxolide. Higher levels were detected for benzophenone (maximum value of 158 ng/g dry weight) and octocrylene (137 ng/g dry weight). In comparison with conventional techniques, this method uses lower amounts of solvents and sorbents, producing less waste ("greener" technique) and comparable performances. In addition, it presents as main advantages the simplicity, speed (short extraction/cleaning time), low cost, and minimum handling of extracts, which can minimize the possibility of samples cross-contamination.

Primary outcome reporting in adolescent depression clinical trials needs standardization.

BACKGROUND: Evidence-based health care is informed by results of randomized clinical trials (RCTs) and their syntheses in meta-analyses. When the trial outcomes measured are not clearly described in trial publications, knowledge synthesis, translation, and decision-making may be impeded. While heterogeneity in outcomes measured in adolescent major depressive disorder (MDD) RCTs has been described, the comprehensiveness of outcome reporting is unknown. This study aimed to assess the reporting of primary outcomes in RCTs evaluating treatments for adolescent MDD. METHODS: RCTs evaluating treatment interventions in adolescents with a diagnosis of MDD published between 2008 and 2017 specifying a single primary outcome were eligible for outcome reporting assessment. Outcome reporting assessment was done independently in duplicate using a comprehensive checklist of 58 reporting items. Primary outcome information provided in each RCT publication was scored as "fully reported", "partially reported", or "not reported" for each checklist item, as applicable. RESULTS: Eighteen of 42 identified articles were found to have a discernable single primary outcome and were included for outcome reporting assessment. Most trials (72%) did not fully report on over half of the 58 checklist items. Items describing masking of outcome assessors, timing and frequency of outcome assessment, and outcome analyses were fully reported in over 70% of trials. Items less frequently reported included outcome measurement instrument properties (ranging from 6 to 17%), justification of timing and frequency of outcome assessment (6%), and justification of criteria used for clinically significant differences (17%). The overall comprehensiveness of reporting appeared stable over time. CONCLUSIONS: Heterogeneous reporting exists in published adolescent MDD RCTs, with frequent omissions of key details about their primary outcomes. These omissions may impair interpretability, replicability, and synthesis of RCTs that inform clinical guidelines and decision-making in this field. Consensus on the minimal criteria for outcome reporting in adolescent MDD RCTs is needed.

Bioconcentration and bioaccumulation of C60 fullerene and C60 epoxide in biofilms and freshwater snails (Radix sp.).

Fullerenes are carbon nanomaterials that have awaken a strong interest due to their adsorption properties and potential applications in many fields. However, there are some gaps of information about their effects and bioconcentration potential in the aquatic biota. In the present work, freshwater biofilms and snails (Radix sp.) were exposed to fullerene C60 aggregates, at concentrations in the low µg/L order, in mesocosms specifically designed to mimic the conditions of a natural stream. The bioconcentration factors of C60 fullerene and its main transformation product, [6,6]C60O epoxide, were studied to the mentioned organisms employing analyses by liquid chromatography coupled to high-resolution mass spectrometry. Our results show that C60 fullerene and its [6,6]C60O present a low bioconcentration factor (BCF) to biofilms: BCFC60 = 1.34 ±â€¯0.95 L/kgdw and BCFC60O = 1.43 ±â€¯0.72 L/kgdw. This suggests that the sorption of these aggregates to biota may be less favoured than it would be suggested by its hydrophobic character. According to our model, the surface of fullerene aggregates is saturated with [6,6]C60O molecules, which exposes the polar epoxide moieties in the surface of the aggregates and decreases their affinity to biofilms. In contrast, freshwater snails showed a moderate capacity to actively retain C60 fullerenes in their organism (BAFC60 = 2670 ±â€¯3070 L/kgdw; BAFC60O = 1330 ±â€¯1680 L/kgdw), probably through ingestion. Our results indicate that the bioaccumulation of these carbon nanomaterials can be hardly estimated using their respective octanol-water partition coefficients, and that their colloidal properties, as well as the feeding strategies of the tested organism, play fundamental roles.

Chronic Kidney Disease Circulating Calciprotein Particles and Extracellular Vesicles Promote Vascular Calcification: A Role for GRP (Gla-Rich Protein).

OBJECTIVE: Inhibition of mineral crystal formation is a crucial step in ectopic calcification. Serum calciprotein particles (CPPs) have been linked to chronic kidney disease (CKD) calcification propensity, but additional knowledge is required to understand their function, assemblage, and composition. The role of other circulating nanostructures, such as extracellular vesicles (EVs) in vascular calcification is currently unknown. Here, we investigated the association of GRP (Gla-rich protein) with circulating CPP and EVs and the role of CKD CPPs and EVs in vascular calcification. APPROACH AND RESULTS: Biological CPPs and EVs were isolated from healthy and CKD patients and comparatively characterized using ultrastructural, analytic, molecular, and immuno-based techniques. Our results show that GRP is a constitutive component of circulating CPPs and EVs. CKD stage 5 serum CPPs and EVs are characterized by lower levels of fetuin-A and GRP, and CPPs CKD stage 5 have increased mineral maturation, resembling secondary CPP particles. Vascular smooth muscle cell calcification assays reveal that CPPs CKD stage 5 and EVs CKD stage 5 are taken up by vascular smooth muscle cells and induce vascular calcification by promoting cell osteochondrogenic differentiation and inflammation. These effects were rescued by incubation of CPPs CKD stage 5 with γ-carboxylated GRP. In vitro, formation and maturation of basic calcium phosphate crystals was highly reduced in the presence of γ-carboxylated GRP, fetuin-A, and MGP (matrix gla protein), and a similar antimineralization system was identified in vivo. CONCLUSIONS: Uremic CPPs and EVs are important players in the mechanisms of widespread calcification in CKD. We propose a major role for cGRP as inhibitory factor to prevent calcification at systemic and tissue levels.

Impact of fullerenes in the bioaccumulation and biotransformation of venlafaxine, diuron and triclosan in river biofilms.

A huge variety of organic microcontaminants are presently detected in freshwater ecosystems, but there is still a lack of knowledge about their interactions, either with living organisms or with other contaminants. Actually, carbon nanomaterials like fullerenes (C60) can act as carriers of organic microcontaminants, but their relevance in processes like bioaccumulation and biotransformation of organic microcontaminants by organisms is unknown. In this study, mesocosm experiments were used to assess the bioaccumulation and biotransformation of three organic microcontaminants (venlafaxine, diuron and triclosan) in river biofilms, and to understand how much the concomitant presence of C60 at environmental relevant concentrations could impact these processes. Results indicated that venlafaxine exhibited the highest bioaccumulation (13% of the initial concentration of venlafaxine in water), while biotransformation was more evident for triclosan (5% of the initial concentration of triclosan in water). Furthermore, biotransformation products such as methyl-triclosan were also present in the biofilm, with levels up to 42% of the concentration of accumulated triclosan. The presence of C60 did not involve relevant changes in the bioaccumulation and biotransformation of microcontaminants in biofilms, which showed similar patterns. Nevertheless, the study shows that a detailed evaluation of the partition of the organic microcontaminants and their transformation products in freshwater systems are important to better understand the impact of the co-existence of others microcontaminants, like carbon nanomaterials, in their possible routes of bioaccumulation and biotransformation.

Synthesis and Biological Evaluation of Ru(II) and Pt(II) Complexes Bearing Carboxyl Groups as Potential Anticancer Targeted Drugs.

The synthesis and characterization of Pt(II) (1 and 2) and Ru(II) arene (3 and 4) or polypyridine (5 and 6) complexes is described. With the aim of having a functional group to form bioconjugates, one uncoordinated carboxyl group has been introduced in all complexes. Some of the complexes were selected for their potential in photodynamic therapy (PDT). The molecular structures of complexes 2 and 5, as well as that of the sodium salt of the 4'-(4-carboxyphenyl)-2,2':6',2″-terpyridine ligand (cptpy), were determined by X-ray diffraction. Different techniques were used to evaluate the binding capacity to model DNA molecules, and MTT cytotoxicity assays were performed against four cell lines. Compounds 3, 4, and 5 showed little tendency to bind to DNA and exhibited poor biological activity. Compound 2 behaves as bonded to DNA probably through a covalent interaction, although its cytotoxicity was very low. Compound 1 and possibly 6, both of which contain a cptpy ligand, were able to intercalate with DNA, but toxicity was not observed for 6. However, compound 1 was active in all cell lines tested. Clonogenic assays and apoptosis induction studies were also performed on the PC-3 line for 1. The photodynamic behavior for complexes 1, 5, and 6 indicated that their nuclease activity was enhanced after irradiation at λ = 447 nm. The cell viability was significantly reduced only in the case of 5. The different behavior in the absence or presence of light makes complex 5 a potential prodrug of interest in PDT. Molecular docking studies followed by molecular dynamics simulations for 1 and the counterpart without the carboxyl group confirmed the experimental data that pointed to an intercalation mechanism. The cytotoxicity of 1 and the potential of 5 in PDT make them good candidates for subsequent conjugation, through the carboxyl group, to "selected peptides" which could facilitate the selective vectorization of the complex toward receptors that are overexpressed in neoplastic cell lines.